A cell-based systems biology assessment of human blood to monitor immune responses after influenza vaccination.
Systems biology is an approach to comprehensively study complex interactions within a biological system. Most published systems vaccinology studies have utilized whole blood or peripheral blood mononuclear cells (PBMC) to monitor the immune response after vaccination. Because human blood is comprised of multiple hematopoietic cell types, the potential for masking responses of under-represented cell populations is increased when analyzing whole blood or PBMC. To investigate the contribution of individual cell types to the immune response after vaccination, we established a rapid and efficient method to purify human T and B cells, natural killer (NK) cells, myeloid dendritic cells (mDC), monocytes, and neutrophils from fresh venous blood. Purified cells were fractionated and processed in a single day. RNA-Seq and quantitative shotgun proteomics were performed to determine expression profiles for each cell type prior to and after inactivated seasonal influenza vaccination. Our results show that transcriptomic and proteomic profiles generated from purified immune cells differ significantly from PBMC. Differential expression analysis for each immune cell type also shows unique transcriptomic and proteomic expression profiles as well as changing biological networks at early time points after vaccination. This cell type-specific information provides a more comprehensive approach to monitor vaccine responses.


Implementing alcohol use disorder pharmacotherapy in primary care settings: a qualitative analysis of provider-identified barriers and impact on implementation outcomes.
Despite the high prevalence of alcohol use disorders (AUDs), in 2016, only 7.8% of individuals meeting diagnostic criteria received any type of AUD treatment. Developing options for treatment within primary care settings is imperative to increase treatment access. As part of a trial to implement AUD pharmacotherapy in primary care settings, this qualitative study analyzed pre-implementation provider interviews using the Consolidated Framework for Implementation Research (CFIR) to identify implementation barriers. Three large Veterans Health Administration facilities participated in the implementation intervention. Local providers were trained to serve as implementation/clinical champions and received external facilitation from the project team. Primary care providers received a dashboard of patients with AUD for case identification, educational materials, and access to consultation from clinical champions. Veterans with AUD diagnoses received educational information in the mail. Prior to the start of implementation activities, 24 primary care providers (5-10 per site) participated in semi-structured interviews. Transcripts were analyzed using common coding techniques for qualitative data using the CFIR codebook Innovation/Intervention Characteristics, Outer Setting, Inner Setting, and Characteristics of Individuals domains. Number and type of barriers identified were compared to quantitative changes in AUD pharmacotherapy prescribing rates. Four major barriers emerged across all three sites: complexity of providing AUD pharmacotherapy in primary care, the limited compatibility of AUD treatment with existing primary care processes, providers' limited knowledge and negative beliefs about AUD pharmacotherapy and providers' negative attitudes toward patients with AUD. Site specific barriers included lack of relative advantage of providing AUD pharmacotherapy in primary care over current practice, complaints about the design quality and packaging of implementation intervention materials, limited priority of addressing AUD in primary care and limited available resources to implement AUD pharmacotherapy in primary care. CFIR constructs were useful for identifying pre-implementation barriers that informed refinements to the implementation intervention. The number and type of pre-implementation barriers identified did not demonstrate a clear relationship to the degree to which sites were able to improve AUD pharmacotherapy prescribing rate. Site-level implementation process factors such as leadership support and provider turn-over likely also interacted with pre-implementation barriers to drive implementation outcomes.


Effects of Light-at-Night on the Rat Liver - A Role for the Autonomic Nervous System.
Exposure to light at night (LAN) has been associated with serious pathologies, including obesity, diabetes and cancer. Recently we showed that 2 h of LAN impaired glucose tolerance in rats. Several studies have suggested that the autonomic nervous system (ANS) plays an important role in communicating these acute effects of LAN to the periphery. Here, we investigated the acute effects of LAN on the liver transcriptome of male Wistar rats. Expression levels of individual genes were not markedly affected by LAN, nevertheless pathway analysis revealed clustered changes in a number of endocrine pathways. Subsequently, we used selective hepatic denervations [sympathetic (Sx), parasympathetic (Px), total (Tx, i.e., Sx plus Px), sham] to investigate the involvement of the ANS in the effects observed. Surgical removal of the sympathetic or parasympathetic hepatic branches of the ANS resulted in many, but small changes in the liver transcriptome, including a pathway involved with circadian clock regulation, but it clearly separated the four denervation groups. On the other hand, analysis of the liver metabolome was not able to separate the denervation groups, and only 6 out of 78 metabolites were significantly up- or downregulated after denervations. Finally, removal of the sympathetic and parasympathetic hepatic nerves combined with LAN exposure clearly modulated the effects of LAN on the liver transcriptome, but left most endocrine pathways unaffected. Conclusion: One-hour light-at-night acutely affects the liver transcriptome. Part of this effect is mediated via the nervous innervation, as a hepatectomy modulated and reduced the effect of LAN on liver transcripts.


Hepatitis C genotype distribution in patient and blood donor samples in South Africa for the period 2008-2012.
There are limited molecular epidemiological studies of hepatitis C at a national level in South Africa. The introduction of newer treatment modalities for hepatitis C requires knowledge of the genotypes as these may have different prognostic and therapeutic implications. This retrospective study describes genotype distributions of patients attending specialist clinics and a blood donor group studied during the period 2008-2012 in South Africa. Residual samples from diagnostic viral load testing from specialist clinics in South Africa (n=941) and from the South African National Blood Service (n=294) were analysed quantitatively by real-time PCR and genotyped using the Versant line probe assay or sequencing. Genotype 1 was predominant in blood donors (34%), whilst genotype 5a was prevalent in patients (36%). In the blood donor group, genotype 4 was detected for the first time. Genotype 2 was rare in the patient group and not detected in blood donors. Genotype 1 was the predominant genotype in the younger age groups (less than 30 years), whereas genotype 5a was found at higher proportions in the older age groups for both the patient and blood donor groups, comprising more than 60% of genotypes in those older than 50 years. Genotypes 1 and 5 were at highest proportions across all provinces compared to other genotypes. In blood donors, genotype 1 was predominant among Caucasians (43%) and genotype 5a among Blacks (54%). Such information is required for planning the impact on the health sector with regard to newly emerging therapies for hepatitis C and burden of disease.


Developmental regulation of full-length trkC in the rat sciatic nerve.
In order to gain insight into potential roles of neurotrophins in Schwann cell biology, the expression of neurotrophin receptors of the trk gene family was investigated in rat sciatic nerve development. This analysis revealed differential regulation of truncated and full-length receptors. TrkA was undetectable even when analysed with a sensitive reverse transcriptase-polymerase chain reaction (RT-PCR) method. TrkB was present at the mRNA as well as protein level only in its truncated form. Surprisingly, multiple isoforms of trkC, including full-length forms, were detected in early postnatal nerve. Specific antibodies detected truncated and full-length trkC proteins in Western blotting, and RT-PCR revealed the presence of two full-length isoforms, one of them containing the 14 amino acid kinase insert. In situ hybridisation localized the expression of trkC to a subpopulation of Schwann cells. TrkC receptors are expressed already in nerves from day-16 embryos. In contrast to early postnatal stages, full-length trkC receptors are no longer expressed in adult nerves, which, however, maintain expression of truncated trkC transcripts. The presence of trkC kinases in peripheral nerve suggests a role for neurotrophin-3, the only known trkC ligand, in peripheral nerve development.


Contribution of astroglia to functionally activated energy metabolism.
Studies of local glucose utilization in neural tissues in vivo with the autoradiographic [14C]deoxyglucose method have demonstrated that energy metabolism increases almost linearly with the degree of functional activation, i.e. spike frequency, in the terminal projection zones of activated pathways. The increased metabolism is found in neuropil and is minimal or undetectable in neuronal cell bodies. Electrical stimulation, increased extracellular [K+] ([K+]o), or opening of Na+ channels with veratridine stimulates metabolism in neutral tissues, and this increase is blocked by ouabain, a specific inhibitor of Na+,K(+)-ATPase. Activation of this enzyme to restore ionic gradients across cellular membranes appears to mediate the function-related increase in energy metabolism. The metabolic activation is, therefore, not directly related to the functional activity itself but to processes operating to recover from that activity. The limited spatial resolution of the [14C]DG method precludes identification of cellular elements in neuropil participating in the metabolic activation, e.g. axonal terminals, dendrites, or astrocytic processes enveloping the synapses. We have, therefore, attempted to stimulate in vitro conditions to be expected from functional activation and increased spike activity in vivo, e.g. increased extracellular [K+], intracellular [Na+], or extracellular neurotransmitter levels, and examined their effects on glucose metabolism in neurons and astroglia in culture. Increased [K+]o stimulated [14C]DG phosphorylation in neuronal and mixed neuronal-astroglial cultures, but not in astroglial cultures assayed in bicarbonate buffer; it did occasionally stimulate metabolism in astroglia when assayed in HEPES or phosphate buffers, but these effects were variable and inconsistent. Veratridine (75 microM) stimulated [14C]DG phosphorylation in neurons and astroglia; these stimulations were blocked by 1 mM ouabain or 10 microM tetrodotoxin (TTX), which blocks voltage-dependent Na+ channels. The Na+ ionophore monensin (10 microM) doubled the rate of metabolism, a stimulation that was only partially blocked by ouabain and unaffected by TTX. L-Glutamate (500 microM) stimulated [14C]DG phosphorylation in astroglia, but this stimulation was probably secondary to Na+ uptake into the cells via a sodium/glutamate co-transporter because it was not blocked by inhibitors of NMDA or non-NMDA receptors but was absent in Na(+)-free medium. These results indicate that astroglia contribute to the increased energy metabolism in neuropil during functional activation by mechanisms that promote Na+ entry into the cells.


Multilocus phylogenies reveal three new truffle-like taxa and the traces of interspecific hybridization in Octaviania (Boletaceae, Boletales).
Among many convergently evolved sequestrate fungal genera in Boletaceae (Boletales, Basidiomycota), the genus Octaviania is the most diverse. We recently collected many specimens of Octaviania subg. Octaviania, including several undescribed taxa, from Japan and the Americas. Here we describe two new species in subgenus Octaviania, O. tenuipes and O. tomentosa, from temperate to subtropical evergreen Fagaceae forests in Japan based on morphological observation and robust multilocus phylogenetic analyses (nrDNA ITS and partial large subunit [LSU], translation elongation factor 1-α gene [TEF1] and the largest subunit of RNA polymerase II gene [RPB1]). Based on specimens from the Americas as well as studies of the holotype, we also taxonomically re-evaluate O. asterosperma var. potteri. Our analysis suggests that O. asterosperma var. potteri is a distinct taxon within the subgenus Octaviania so we recognize this as O. potteri stat. nov. We unexpectedly collected O. potteri specimens from geographically widespread sites in the USA, Japan and Colombia. This is the first verified report of Octaviania from the South American continent. Our molecular analyses also revealed that the RPB1 sequence of one O. tenuipes specimen was identical to that of a closely related species, O. japonimontana, and that one O. potteri specimen from Minnesota had an RPB1 sequence of an unknown species of O. subg. Octaviania. Additionally, one O. japonimontana specimen had an unusually divergent TEF1 sequence. Gene-tree comparison and phylogenetic network analysis of the multilocus dataset suggest that these heterogenous sequences are most likely the result of previous inter- and intra-specific hybridization. We hypothesize that frequent hybridization events in Octaviania may have promoted the high genetic and species diversity found within the genus.


Decoding Articulatory Features from fMRI Responses in Dorsal Speech Regions.
The brain's circuitry for perceiving and producing speech may show a notable level of overlap that is crucial for normal development and behavior. The extent to which sensorimotor integration plays a role in speech perception remains highly controversial, however. Methodological constraints related to experimental designs and analysis methods have so far prevented the disentanglement of neural responses to acoustic versus articulatory speech features. Using a passive listening paradigm and multivariate decoding of single-trial fMRI responses to spoken syllables, we investigated brain-based generalization of articulatory features (place and manner of articulation, and voicing) beyond their acoustic (surface) form in adult human listeners. For example, we trained a classifier to discriminate place of articulation within stop syllables (e.g., /pa/ vs /ta/) and tested whether this training generalizes to fricatives (e.g., /fa/ vs /sa/). This novel approach revealed generalization of place and manner of articulation at multiple cortical levels within the dorsal auditory pathway, including auditory, sensorimotor, motor, and somatosensory regions, suggesting the representation of sensorimotor information. Additionally, generalization of voicing included the right anterior superior temporal sulcus associated with the perception of human voices as well as somatosensory regions bilaterally. Our findings highlight the close connection between brain systems for speech perception and production, and in particular, indicate the availability of articulatory codes during passive speech perception. Sensorimotor integration is central to verbal communication and provides a link between auditory signals of speech perception and motor programs of speech production. It remains highly controversial, however, to what extent the brain's speech perception system actively uses articulatory (motor), in addition to acoustic/phonetic, representations. In this study, we examine the role of articulatory representations during passive listening using carefully controlled stimuli (spoken syllables) in combination with multivariate fMRI decoding. Our approach enabled us to disentangle brain responses to acoustic and articulatory speech properties. In particular, it revealed articulatory-specific brain responses of speech at multiple cortical levels, including auditory, sensorimotor, and motor regions, suggesting the representation of sensorimotor information during passive speech perception.


General solutions to consistency problems in multiple hypothesis testing.
Coherence and consonance are the two important concepts to ensure consistent conclusions drawn from multiple tests. Unlike coherence, consonance of a multiple testing procedure (MTP) has not yet received much attention. Although most of the MTPs used in practice are consonant, cases still exist that dissonant tests have to be used due to their unique advantages, for example, the likelihood ratio tests and sum tests. Consonance adjustments are necessary for such tests to avoid difficulty in interpretation. Moreover, in terms of detecting elementary hypotheses, a consonant test procedure is uniformly more powerful than the corresponding dissonant one. In this paper, several general methods using the partitioning principle to construct (strongly) consonant and strongly coherent MTPs are proposed. As a result, any dissonant (but coherent) multiple level α test can be improved by a consonant one without any cost in terms of inferences on elementary hypotheses.


Comparing an Emotion- and a Behavior-Focused Parenting Program as Part of a Multsystemic Intervention for Child Conduct Problems.
This study evaluated the effectiveness of a multisystemic early intervention that included a comparison of an emotion- and behavior-focused parenting program for children with emerging conduct problems. The processes that moderated positive child outcomes were also explored. A repeated measures cluster randomized group design methodology was employed with three conditions (Tuning in to Kids, Positive Parenting Program, and waitlist control) and two periods (preintervention and 6-month follow-up). The sample consisted of 320 predominantly Caucasian 4- to 9-year-old children who were screened for disruptive behavior problems. Three outcome measures of child conduct problems were evaluated using a parent (Eyberg Child Behavior Inventory) and teacher (Strengths and Difficulties Questionnaire) rating scale and a structured child interview (Home Interview With Child). Six moderators were assessed using family demographic information and a parent-rated measure of psychological well-being (Depression Anxiety and Stress Scales short form). The results indicated that the multisystemic intervention was effective compared to a control group and that, despite different theoretical orientations, the emotion- and behavior-focused parenting programs were equally effective in reducing child conduct problems. Child age and parent psychological well-being moderated intervention response. This effectiveness trial supports the use of either emotion- or behavior-focused parenting programs in a multisystemic early intervention and provides greater choice for practitioners in the selection of specific programs.


Effect of leukocyte compatibility on neutrophil increment after transfusion of granulocyte colony-stimulating factor-mobilized prophylactic granulocyte transfusions and on clinical outcomes after stem cell transplantation.
The primary limitations of granulocyte transfusions include low component cell dose and leukocyte incompatibility. Component cell dose improved with granulocyte colony-stimulating factor (G-CSF) mobilization, and the transfusion of G-CSF-mobilized, human leukocyte antigen (HLA)-matched granulocyte components resulted in significant, sustained absolute neutrophil count (ANC) increments. However, the effect of leukocyte compatibility on outcomes with G-CSF-mobilized granulocyte transfusions is unclear. The objectives were to determine the effect of leukocyte compatibility on ANC increments and selected clinical outcomes after transfusion of prophylactic, G-CSF-mobilized granulocyte components into neutropenic recipients of autologous peripheral blood stem cell (PBSC) transplants. Beginning on transplant day 2, 23 evaluable recipients were scheduled to receive 4 alternate-day transfusions of granulocyte components apheresed from a single donor given G-CSF. G-CSF was also given to recipients after transplantation. Recipient ANC was determined before and sequentially after each granulocyte transfusion to determine the peak ANC increment. Leukocyte compatibility was determined at study entry only by a lymphocytotoxicity screening assay (s-LCA) against a panel of HLA-defined cells. Eight recipients had positive s-LCA. On days 2 and 4, the mean peak ANC increments after granulocyte transfusion were comparable between the cohorts with positive and negative s-LCA. However, the mean peak ANC increments on day 6 (246/microL vs 724/microL; P =.05) and day 8 (283/microL vs 1079/microL; P =.06) were lower in the cohort with positive s-LCA, in spite of the transfusion of comparable component cell doses. Adverse reactions occurred with only 5 of 87 (5.7%) granulocyte transfusions and were not associated with leukocyte compatibility test results. Platelet increments, determined 1 hour after granulocyte transfusion, were comparable between the cohorts. Although the 2 cohorts received PBSC components with similar CD34(+) cell doses, the cohort with a positive s-LCA had delayed neutrophil engraftment and a greater number of febrile days and required more days of intravenous antibiotics and platelet transfusions. Leukocyte incompatibility adversely affected ANC increments after the transfusion of G-CSF-mobilized granulocyte components and clinical outcomes after PBSC transplantation.


Child behavior and accidental injury in 11,966 preschool children.
Social and behavioral characteristics of 11,966 British children, aged 5 years, and mothers' reports of accidental injuries between birth and age 5 years were analyzed. Aggressive behavior was associated with all accidental injuries after controlling psychosocial variables including social class; crowding; mother's psychological distress, age, and marital status; and child's sex. Overactivity was associated only with injuries not resulting in hospitalization after control of the covariates. The relative risk of injuries resulting in hospitalization was 1.9 among children with both high activity and high aggression scores compared with children with low scores on both behavioral scales. The findings support the inference that aggression and overactivity are independently associated with accidents. The associations between child behavior and injuries were stronger than the associations between injuries and the social factors including social class and crowding. This finding suggests that interventions aimed at high-risk groups may be effective supplements to environmental interventions.


Influence of ketamine and propranolol on plasma renin activity (PRA) in female rabbits and guinea pigs.
1. Plasma renin activity was measured in non pregnant rabbits and guinea pigs under Ketamine-induced general anesthesia after pretreatment either with Propranolol or with a Placebo. Study was performed using a radio-immunoassay for angiotensin I. 2. Twenty minutes after the beginning of the observation period, renin activity in rabbits who had received Placebo alone (11.47 +/- 2.35 ng/ml/h) or associated with Ketamine (11.36 +/- 2.54 ng/ml/h) was similar. However, enzyme activity was significantly lower (P less than 0.001) when Propranolol was associated with Ketamine (3.97 +/- 0.58 ng/ml/h) or with Placebo (4.10 +/- 0.55 ng/ml/h). 3. In the same way, renin activity was significantly higher (P less than 0.001) in guinea pigs without Propranolol than in those who had received this drug. 4. These findings indicate that stress induced by general anesthesia with Ketamine or by simple manipulation of animals (Placebo) was accompanied by an excessive increase in plasma renin activity. Propranolol maintained the level of this enzyme activity within normal limits.


Monolith peptide affinity chromatography for quantification of immunoglobulin M.
We have developed a method for quantification of a specific monoclonal IgM directed toward embryonic stem cells based on a peptide affinity monolith. A peptide affinity ligand with the sequence C-C-H-Q-R-L-S-Q-R-K was obtained by epitope mapping using peptide SPOT synthesis. The peptide ligand was covalently immobilized by coupling the N-terminal cysteine to a monolithic disk that was previously modified with iodated spacer molecules. The monolithic disc was used for quantification of purified IgM and for IgM present in mammalian cell culture supernatant. We observed 17% unspecific binding of IgM to the monolithic disk and additionally a product loss in the flow through of 20%. Nevertheless, calibration curves had high correlation coefficients and inter/intra-assay variability experiments proved sufficient precision of the method. A limit of quantification of 51.69 μg/mL for purified IgM and 48.40 μg/mL for IgM in cell culture supernatant could be calculated. The binding capacity was consistent within the period of the study which included more than 200 cycles. The analysis time of less than 2 min is an advantage over existing chromatographic methods that rely on pore diffusion.


Diagnosis in vascular dementia, applying 'Cochrane diagnosis rules' to 'dementia diagnostic tools'.
In this issue of Clinical Science, Biesbroek and colleagues describe recent work on magnetic resonance imaging (MRI)-based cerebral lesion location and its association with cognitive decline. The authors conclude that diagnostic neuroimaging in dementia should shift from whole-brain evaluation to focused quantitative analysis of strategic brain areas. This commentary uses the review of lesion location mapping to discuss broader issues around studies of dementia test strategies. We draw upon work completed by the Cochrane Dementia and Cognitive Improvement Group designed to improve design, conduct and reporting of dementia biomarker studies.


Exercise training in patients with advanced chronic heart failure (NYHA IIIb) promotes restoration of peripheral vasomotor function, induction of endogenous regeneration, and improvement of left ventricular function.
Attenuated peripheral perfusion in patients with advanced chronic heart failure (CHF) is partially the result of endothelial dysfunction. This has been causally linked to an impaired endogenous regenerative capacity of circulating progenitor cells (CPC). The aim of this study was to elucidate whether exercise training (ET) affects exercise intolerance and left ventricular (LV) performance in patients with advanced CHF (New York Heart Association class IIIb) and whether this is associated with correction of peripheral vasomotion and induction of endogenous regeneration. Thirty-seven patients with CHF (LV ejection fraction 24+/-2%) were randomly assigned to 12 weeks of ET or sedentary lifestyle (control). At the beginning of the study and after 12 weeks, maximal oxygen consumption (Vo(2)max) and LV ejection fraction were determined; the number of CD34(+)/KDR(+) CPCs was quantified by flow cytometry and CPC functional capacity was determined by migration assay. Flow-mediated dilation was assessed by ultrasound. Capillary density was measured in skeletal muscle tissue samples. In advanced CHF, ET improved Vo(2)max by +2.7+/-2.2 versus -0.8+/-3.1 mL/min/kg in control (P=0.009) and LV ejection fraction by +9.4+/-6.1 versus -0.8+/-5.2% in control (P<0.001). Flow-mediated dilation improved by +7.43+/-2.28 versus +0.09+/-2.18% in control (P<0.001). ET increased the number of CPC by +83+/-60 versus -6+/-109 cells/mL in control (P=0.014) and their migratory capacity by +224+/-263 versus -12+/-159 CPC/1000 plated CPC in control (P=0.03). Skeletal muscle capillary density increased by +0.22+/-0.10 versus -0.02+/-0.16 capillaries per fiber in control (P<0.001). Twelve weeks of ET in patients with advanced CHF is associated with augmented regenerative capacity of CPCs, enhanced flow-mediated dilation suggestive of improvement in endothelial function, skeletal muscle neovascularization, and improved LV function. Clinical Trial Registration- http://www.clinicaltrials.gov. Unique Identifier: NCT00176384.


An open label pilot study of a dexmedetomidine-remifentanil-caudal anesthetic for infant lower abdominal/lower extremity surgery: The T REX pilot study.
Concern over potential neurotoxicity of anesthetics has led to growing interest in prospective clinical trials using potentially less toxic anesthetic regimens, especially for prolonged anesthesia in infants. Preclinical studies suggest that dexmedetomidine may have a reduced neurotoxic profile compared to other conventional anesthetic regimens; however, coadministration with either anesthetic drugs (eg, remifentanil) and/or regional blockade is required to achieve adequate anesthesia for surgery. The feasibility of this pharmacological approach is unknown. The aim of this study was to determine the feasibility of a remifentanil/dexmedetomidine/neuraxial block technique in infants scheduled for surgery lasting longer than 2 hours. Sixty infants (age 1-12 months) were enrolled at seven centers over 18 months. A caudal local anesthetic block was placed after induction of anesthesia with sevoflurane. Next, an infusion of dexmedetomidine and remifentanil commenced, and the sevoflurane was discontinued. Three different protocols with escalating doses of dexmedetomidine and remifentanil were used. One infant was excluded due to a protocol violation and consent was withdrawn prior to anesthesia in another. The caudal block was unsuccessful in two infants. Of the 56 infants who completed the protocol, 45 (80%) had at least one episode of hypertension (mean arterial pressure >80 mm Hg) and/or movement that required adjusting the anesthesia regimen. In the majority of these cases, the remifentanil and/or dexmedetomidine doses were increased although six infants required rescue 0.3% sevoflurane and one required a propofol bolus. Ten infants had at least one episode of mild hypotension (mean arterial pressure 40-50 mm Hg) and four had at least one episode of moderate hypotension (mean arterial pressure <40 mm Hg). A dexmedetomidine/remifentanil neuraxial anesthetic regimen was effective in 87.5% of infants. These findings can be used as a foundation for designing larger trials that assess alternative anesthetic regimens for anesthetic neurotoxicity in infants.


Anticholinergics for symptomatic management of Parkinson's disease.
Anticholinergics were the first drugs available for the symptomatic treatment of Parkinson's disease and they are still widely used today, both as monotherapy and as part of combination regimes. They are commonly believed to be associated with a less favourable side effect profile than other antiparkinsonian drugs, in particular with respect to neuropsychiatric and cognitive adverse events. They have been claimed to exert a better effect on tremor than on other parkinsonian features. To determine the efficacy and tolerability of anticholinergics in the symptomatic treatment of Parkinson's disease compared to placebo or no treatment. The literature search included electronic searches of the Cochrane Controlled Trials Register (The Cochrane Library, Issue 4, 2001), MEDLINE (1966 to 2001), Old Medline (1960-1965), Index Medicus (1927 - 1959), as well as handsearching the neurology literature including the reference lists of identified articles, other reviews and book chapters. Randomised controlled trials of anticholinergic drugs versus placebo or no treatment in de-novo or advanced Parkinson's disease, either as monotherapy or as an add-on to other antiparkinsonian drugs were included. Trials of anticholinergic drugs that were never in general clinical use were excluded. Data was abstracted independently by two authors. Differences were settled by discussion among all authors. Data collected included patient characteristics, disease duration and severity, concomitant medication, interventions including duration and dose of anticholinergic treatment, outcome measures, rates of and reasons for withdrawals, and neuropsychiatric and cognitive adverse events. The initial search yielded 14 potentially eligible studies, five of which were subsequently excluded. In three cases this was because they dealt with substances that had never been marketed or had not been licensed for as far as could be traced back. One trial had been published twice in different languages. One study was excluded based on the assessment of its methodological quality. The remaining nine studies were all of double-blind cross-over design and included 221 patients. Trial duration was between five and 20 weeks and drugs investigated were benzhexol (mean doses: 8 to 20 mg/d), orphenadrine (mean dose not reported), benztropine (mean dose not reported), bornaprine (8 to 8.25 mg/d), benapryzine (200 mg/d), and methixine (45 mg/d). Only one study involved two anticholinergic drugs. Outcome measures varied widely across studies and in many cases, the scales applied were the authors' own and were not defined in detail. Incomplete reporting of methodology and results was frequent. The heterogeneous study designs as well as incomplete reporting precluded combined statistical analysis. Five studies used both tremor and other parkinsonian features as outcome measures. Outcome measures in these five studies were too different for a combined analysis and results varied widely, from a significant improvement in tremor only to significant improvement in other features but not in tremor. All studies except one (dealing with methixine) found a significant improvement from baseline on the anticholinergic drug in at least one outcome measure. The difference between placebo and active drug was reported in four studies and was found to be significant in all cases. No study failed to show superiority of the anticholinergic over placebo. The occurrence of neuropsychiatric and cognitive adverse events was reported in all but three studies (in 35 patients on active drug versus 13 on placebo). The most frequently reported reason for drop-outs from studies was in patients on placebo due to withdrawal from pre-trial anticholinergic treatment. As monotherapy or as an adjunct to other antiparkinsonian drugs, anticholinergics are more effective than placebo in improving motor function in Parkinson's disease. Neuropsychiatric and cognitive adverse events occur more frequently on anticholinergics than on placebo and are a more common reason for withdrawal than lack of efficacy. Results regarding a potentially better effect of the anticholinergic drug on tremor than on other outcome measures are conflicting and data do not strongly support a differential clinical effect on individual parkinsonian features. Data is insufficient to allow comparisons in efficacy or tolerability between individual anticholinergic drugs.


Literature based drug interaction prediction with clinical assessment using electronic medical records: novel myopathy associated drug interactions.
Drug-drug interactions (DDIs) are a common cause of adverse drug events. In this paper, we combined a literature discovery approach with analysis of a large electronic medical record database method to predict and evaluate novel DDIs. We predicted an initial set of 13197 potential DDIs based on substrates and inhibitors of cytochrome P450 (CYP) metabolism enzymes identified from published in vitro pharmacology experiments. Using a clinical repository of over 800,000 patients, we narrowed this theoretical set of DDIs to 3670 drug pairs actually taken by patients. Finally, we sought to identify novel combinations that synergistically increased the risk of myopathy. Five pairs were identified with their p-values less than 1E-06: loratadine and simvastatin (relative risk or RR = 1.69); loratadine and alprazolam (RR = 1.86); loratadine and duloxetine (RR = 1.94); loratadine and ropinirole (RR = 3.21); and promethazine and tegaserod (RR = 3.00). When taken together, each drug pair showed a significantly increased risk of myopathy when compared to the expected additive myopathy risk from taking either of the drugs alone. Based on additional literature data on in vitro drug metabolism and inhibition potency, loratadine and simvastatin and tegaserod and promethazine were predicted to have a strong DDI through the CYP3A4 and CYP2D6 enzymes, respectively. This new translational biomedical informatics approach supports not only detection of new clinically significant DDI signals, but also evaluation of their potential molecular mechanisms.


MDS-based multiresolution nonlinear dimensionality reduction model for color image segmentation.
In this paper, we present an efficient coarse-to-fine multiresolution framework for multidimensional scaling and demonstrate its performance on a large-scale nonlinear dimensionality reduction and embedding problem in a texture feature extraction step for the unsupervised image segmentation problem. We demonstrate both the efficiency of our multiresolution algorithm and its real interest to learn a nonlinear low-dimensional representation of the texture feature set of an image which can then subsequently be exploited in a simple clustering-based segmentation algorithm. The resulting segmentation procedure has been successfully applied on the Berkeley image database, demonstrating its efficiency compared to the best existing state-of-the-art segmentation methods recently proposed in the literature.


Two cases of dysuria as a complication of single-session bilateral triple pelvic osteotomy.
Dysuria is a rare, but nevertheless severe postoperative complication associated with single-session bilateral triple pelvic osteotomy. Two cases are described in which contrast radiography of the lower urinary tract was used to diagnose urethral compression due to pelvic canal narrowing. Relief of the pressure on the urethra was achieved with an external fixation device, which distracted the acetabular fragments of the pelvis. In both cases the dysuria was resolved. The significance of this complication and its possible causes, especially regarding surgical technique, are discussed.


A randomized trial assessing the impact of written information on outpatients' knowledge about and attitude toward randomized clinical trials. The INFO trial group.
To improve the patient education process in clinical research, three information materials describing general aspects of design and conduct of randomized clinical trials were developed. The materials varied in length, reading ability level, and reader appeal. Their influence on knowledge about and attitude toward randomized clinical trials was assessed in a randomized, parallel group, evaluator-blinded trial among 415 outpatients. The patients were randomized to the following groups: control (no intervention), leaflet, brochure, or booklet. Knowledge was assessed by a 17-item multiple-choice questionnaire and attitude was assessed by a 32-item Likert questionnaire at entry and 2 weeks after the intervention. The interventions and the questionnaires were pilot tested and power calculations were performed. At entry, the mean knowledge score was 7.9 points. At follow-up, the knowledge scores increased by 0.5 for the control, 1.0 for the leaflet, 1.6 for the brochure, and 1.4 for the booklet. The brochure and the booklet improved the knowledge score significantly compared with the control. The general attitude was positive at entry (mean 71.5 points). Only the booklet significantly increased the total attitude score (4.8 points) and the randomized clinical trials attitude subscale score (1.8 points). In conclusion, written information significantly improved outpatients' knowledge about and attitude toward randomized clinical trials. Detailed rather than brief information was more effective. Control Clin Trials 2000;21:223-240


Coffee Consumption and Risk of Gallbladder Cancer in a Prospective Study.
Evidence indicates that coffee consumption may reduce the risk of gallstone disease, which is strongly associated with increased risk of gallbladder cancer. The association between coffee consumption and gallbladder cancer incidence was examined in a prospective cohort study of 72 680 Swedish adults (aged 45 - 83 years) who were free of cancer and reported their coffee consumption at baseline. Gallbladder cancers were ascertained by linkage with the Swedish Cancer Register. The data were analyzed using Cox proportional hazards regression models. Statistical tests were two-sided. During 967 377 person-years of follow-up, 74 gallbladder cancer case patients were identified. Compared with consumption of one or less cups of coffee per day, the multivariable hazard ratios were 0.76 (95% confidence interval [CI] = 0.41 to 1.41) for two cups per day, 0.50 (95% CI = 0.24 to 1.06) for three cups per day, and 0.41 (95% CI = 0.20 to 0.83) for four or more cups per day. In conclusion, coffee consumption is associated with a reduced risk of gallbladder cancer.


Disruption of the mouse xeroderma pigmentosum group D DNA repair/basal transcription gene results in preimplantation lethality.
The xeroderma pigmentosum (XP) group D (XPD) gene encodes a DNA helicase that is a subunit of the transcription factor IIH complex, involved both in nucleotide excision repair of UV-induced DNA damage and in basal transcription initiation. Point mutations in the XPD gene lead either to the cancer-prone repair syndrome XP, sometimes in combination with a second repair condition; Cockayne syndrome; or the non-cancer-prone brittle-hair disorder trichothiodystrophy. To study the role of XPD in nucleotide excision repair and transcription and its implication in human disorders, we isolated the mouse XPD gene and generated a null allele via homologous recombination in embryonic stem cells by deleting XPD helicase domains IV-VI. Heterozygous cells and mice are normal without any obvious defect. However, when intercrossing heterozygotes, homozygous XPD mutant mice were selectively absent from the offspring. Furthermore, we could not detect XPD-/- embryos at day 7.5 of development. In vitro growth experiments with preimplantation-stage embryos obtained from heterozygous intercrosses showed a significantly higher fraction of embryos that died at the two-cell stage, compared to wild-type embryos. These results establish the essential function of the XPD protein in mammals and in cellular viability and are consistent with the notion that only subtle XPD mutations are found in XP, XP/Cockayne syndrome, and trichothiodystrophy patients.


Comparison of immune responses in patients infected with Vibrio cholerae O139 and O1.
Vibrio cholerae O139 has recently emerged as the second etiologic agent of cholera in Asia. A study was carried out to evaluate the induction of specific immune responses to the organism in V. cholerae O139-infected patients. The immune responses to V. cholerae O139 Bengal were studied in patients by measuring antibody-secreting cells (ASC), as well as vibriocidal and antitoxic antibodies in the circulation. These responses were compared with those in patients with V. cholerae O1 disease. Strong immunoglobulin A (IgA) and IgM ASC responses were seen against the homologous lipopolysaccharide or serogroup of V. cholerae. The magnitude and isotype of the responses were similar in O139- and O1-infected patients. Vibriocidal antibody responses were seen against bacteria of the homologous but not heterologous serogroup, and these responses reflect the lack of cross-protection between the infections caused by the two serogroups. The two groups of patients showed comparable cholera toxin-specific ASC responses, with the IgG isotype dominating over the IgA isotype, as well as comparable antitoxic immune responses in plasma. These results suggest that despite having a polysaccharide capsule, V. cholerae O139 induces systemic and intestine-derived ASC responses in peripheral blood comparable to those seen in patients with V. cholerae O1 disease.


The effect of free personal care for the elderly on informal caregiving.
Population forecasters have predicted that the proportion of people in the UK aged 65 years and older will rise significantly in coming decades. This shift in demographics will put increasing pressure on the National Health Service and providers of social care. However, older people do not rely only on care provided by the state; informal care of the elderly is often supplied by family and friends. Therefore, the relationship between formal and informal care and the reaction of informal carers to institutional changes is an important policy issue. This study uses individual level data from the British Household Panel Survey to estimate the effects of the introduction of free personal care for the elderly in Scotland on informal care behaviour. As the change in policy applied only to Scotland, a natural experiment is formed allowing a difference-in-differences approach to be used. This paper finds that the introduction of the policy increased the probability of women supplying informal care by around six percentage points. In addition, for both sexes, it reports evidence of a shift away from the upper and lower tails towards the middle of the hours of care distribution as a result of the change in policy.


[Epidemiology, clinical spectrum of ALS and differential diagnoses].
Amyotrophic Lateral Sclerosis (ALS) is the most common motor neuron disease in adults. Its incidence in France is estimated at 2.5 per 100,000 population and its prevalence between 5 and 8 per 100,000 inhabitants. Good prognostic factors are age of early onset, a longer time to diagnosis, initial damage to the spinal onset, early management of undernutrition and restrictive respiratory failure. The diagnosis of ALS is primarily clinical and is based on the evidence of involvement of the central motor neuron and peripheral neuron (NMP) in different territories or spinal or bulbar. The EMG confirms the achievement of NMP, shows the extension to clinically preserved areas and allows to exclude some differential diagnoses. The clinical spectrum of ALS is broad: conventional forms beginning brachial, lower limb or bulbar onsets, rarer forms to start breathing, pyramidal forms, forms with cognitive and behavioural impairment. In 5-10% of cases, ALS is familial. In 15% of cases, it is associated with frontotemporal degeneration rather than orbito-frontal type. The main differential diagnoses are guided by the clinic: combining pure motor neuropathy with or without conduction block, post-polio syndrome, cramp-fasciculation syndrome, myasthenia gravis, paraneoplastic syndromes, Sjögren syndrome, retroviral infections, some endocrine disorders, some metabolic diseases, genetic diseases (Kennedy and SMA) and inclusion body myositis.


Hindlimb patterning and mandible development require the Ptx1 gene.
The restricted expression of the Ptx1 (Pitx1) gene in the posterior half of the lateral plate mesoderm has suggested that it may play a role in specification of posterior structures, in particular, specification of hindlimb identity. Ptx1 is also expressed in the most anterior ectoderm, the stomodeum, and in the first branchial arch. Ptx1 expression overlaps with that of Ptx2 in stomodeum and in posterior left lateral plate mesoderm. We now show that targeted inactivation of the mouse Ptx1 gene severely impairs hindlimb development: the ilium and knee cartilage are absent and the long bones are underdeveloped. Greater reduction of the right femur size in Ptx1 null mice suggests partial compensation by Ptx2 on the left side. The similarly sized tibia and fibula of mutant hindlimbs may be taken to resemble forelimb bones: however, the mutant limb buds appear to have retained their molecular identity as assessed by forelimb expression of Tbx5 and by hindlimb expression of Tbx4, even though Tbx4 expression is decreased in Ptx1 null mice. The hindlimb defects appear to be, at least partly, due to abnormal chondrogenesis. Since the most affected structures derive from the dorsal side of hindlimb buds, the data suggest that Ptx1 is responsible for patterning of these dorsal structures and that as such it may control development of hindlimb-specific features. Ptx1 inactivation also leads to loss of bones derived from the proximal part of the mandibular mesenchyme. The dual role of Ptx1 revealed by the gene knockout may reflect features of the mammalian jaw and hindlimbs that were acquired at a similar time during tetrapod evolution.


Allium vegetable consumption and health: An umbrella review of meta-analyses of multiple health outcomes.
Previously, our meta-analysis and other studies have suggested that allium vegetable consumption is beneficial for health, but no umbrella review has been conducted to assess the evidence of the various health benefits of allium vegetable consumption. Therefore, we conducted this umbrella review on this topic. This umbrella review included a total of 16 meta-analyses with 50 unique outcomes. The most beneficial cancer-related outcome was shown for gastric cancer (risk ratio 0.78; 95% confidence interval [CI] 0.67-0.91). In addition, only 8 weeks of garlic consumption significantly decreased serum total cholesterol (weighted mean differences -17.20 mg/dl; 95% CI -23.10 to -11.30), and patients with dyslipidemia who consumed garlic experienced more benefits than the whole population. Diabetic patients with longer durations of garlic intake experienced more benefits in terms of fasting blood glucose (FBG), HbA1c, and serum fructosamine than healthy participants, and garlic intake was associated with blood pressure reduction in hypertensive patients but not in normotensive participants. Limited side effects of garlic, such as garlic odor and gastrointestinal complaints, were reported among the included meta-analyses. Our results suggested that allium vegetables might be beneficial for cancer prevention. In particular, garlic was comparatively safe and is recommended as a long-term dietary component for patients with dyslipidemia, diabetes, and hypertension.


Influence of late-Holocene climate change on the solid-phase speciation and long-term stability of arsenic in sub-Arctic lake sediments.
Sediment cores were collected from two lakes in the Courageous Lake Greenstone Belt (CLGB), central Northwest Territories, Canada, to examine the influence of late-Holocene warming on the transport and fate of arsenic (As) in sub-Arctic lakes. In both lakes, allochthonous As-bearing minerals (i.e. arsenopyrite and scorodite) were identified in sediment deposited during times of both regional warming and cooling, suggesting that weathering of bedrock and derived surficial materials provides a continual source of As to lakes of the CLGB. However, maximum porewater As (84 μg·L-1 and 15 μg·L-1) and reactive organic matter (OM; aquatic and terrestrial-derived) concentrations in each lake are coincident with known periods of regional climate warming. It is inferred that increased biological production in surface waters and influx of terrigenous OM led to the release of sedimentary As to porewater through reductive dissolution of As-bearing Fe-(oxy)hydroxides and scorodite during episodes of regional warming. Elevated sedimentary As concentrations (median: 36 mg·kg-1; range: 29 to 49 mg·kg-1) are observed in sediment coeval with the Holocene Thermal Maximum (ca. 5430 ± 110 to 4070 ± 130 cal. years BP); at these depths, authigenic As-bearing framboidal pyrite is the primary host of As in sediment and the influence of organic matter on the precipitation of As-bearing framboidal pyrite is apparent petrographically. These findings suggest that increased biological productivity and weathering of terrestrial OM associated with climate warming influences redox cycles in the near-surface sediment and enhances the mobility of As in northern lakes. Knowledge generated from this study is relevant for predicting future climate change-driven variations in metal(loid) cycling in aquatic systems and can be used to interpret trends in long-term environmental monitoring data at historical, modern, and future metal mines in northern environments.


Trends and Most Frequent Methods of Suicide in Chile Between 2001 and 2010.
Despite the many studies trying to evaluate the magnitude of suicide in Chile, none of them include the new valid data, recently published by the DEIS of the Chilean Health Ministry. This paper sought to describe how suicide rates changed among Chileans who were at least 15 years of age during the period of 2001-2010; these rates were stratified by sex, age, urban/rural status, and region of the country. An exploratory and temporal study was conducted. Suicide mortality rates were calculated by collecting information from the Chilean Ministry of Health's death registry between 2001 and 2010 among subjects who were at least 15 years of age. Crude and age-adjusted mortality rates were computed. The standardized suicide rate in Chile during 2001-2010 was 14.7 deaths per 100,000, the highest in South America. The minimum age-adjusted suicide rate observed during the examined period was 13.49 per 100,000 in 2005, and the maximum, 16.67 per 100,000 in 2008. Suicide rates among men were up to 4.8 times the suicide rates among women. In addition, rates in rural areas were double compared with urban areas. Hanging was the most common suicide method, followed by use of firearms and explosives in men and poisoning in women. Finally, there was a progressive increase in suicide rates in the southernmost regions of the country. Suicide rates in Chile are relatively high among male subjects in rural areas. There is a disturbing increase in suicide rates among younger individuals and women, although these rates remain lower among younger individuals and women than among adults in general.


Quality control of leather by terahertz time-domain spectroscopy.
We use terahertz time-domain spectroscopy, combined with effective-medium theory, to measure the moisture content and thickness of leather simultaneously. These results demonstrate that this method could become a standard quality control test for the industrial tanning process.


Kleptoplastidic benthic foraminifera from aphotic habitats: insights into assimilation of inorganic C, N and S studied with sub-cellular resolution.
The assimilation of inorganic compounds in foraminiferal metabolism compared to predation or organic matter assimilation is unknown. Here, we investigate possible inorganic-compound assimilation in Nonionellina labradorica, a common kleptoplastidic benthic foraminifer from Arctic and North Atlantic sublittoral regions. The objectives were to identify the source of the foraminiferal kleptoplasts, assess their photosynthetic functionality in light and darkness and investigate inorganic nitrogen and sulfate assimilation. We used DNA barcoding of a ~ 830 bp fragment from the SSU rDNA to identify the kleptoplasts and correlated transmission electron microscopy and nanometre-scale secondary ion mass spectrometry (TEM-NanoSIMS) isotopic imaging to study 13 C-bicarbonate, 15 N-ammonium and 34 S-sulfate uptake. In addition, respiration rate measurements were determined to assess the response of N. labradorica to light. The DNA sequences established that over 80% of the kleptoplasts belonged to Thalassiosira (with 96%-99% identity), a cosmopolitan planktonic diatom. TEM-NanoSIMS imaging revealed degraded cytoplasm and an absence of 13 C assimilation in foraminifera exposed to light. Oxygen measurements showed higher respiration rates under light than dark conditions, and no O2 production was detected. These results indicate that the photosynthetic pathways in N. labradorica are not functional. Furthermore, N. labradorica assimilated both 15 N-ammonium and 34 S-sulfate into its cytoplasm, which suggests that foraminifera might have several ammonium or sulfate assimilation pathways, involving either the kleptoplasts or bona fide foraminiferal pathway(s) not yet identified.


Clinical features of confirmed versus suspected urogenital tuberculosis in region with extremely high prevalence of pulmonary tuberculosis.
To compare the characteristics of confirmed vs suspected cases of urogenital tuberculosis (UGTB) in a geographic region with an extremely high prevalence of pulmonary tuberculosis. UGTB is notoriously difficult to diagnose. A retrospective clinical record review was performed of 68 patients treated from March 1998 to July 2007. Group 1 (n = 45) had UGTB confirmed by microbiologic or histologic examination. Group 2 (n = 23) had a high suspicion of UGTB because of the clinical features, but no microbiologic or histologic confirmation. The data were collected and statistically analyzed using Student's t test for parametric data and Fisher's exact test for contingency tables (P < .05 was accepted as statistically significant). The clinical characteristics were not significantly different statistically, except for flank pain (14% vs 43%), renal cavitation (14% vs 44%), urolithiasis (0% vs 25%), and ureteral stricture formation (7% vs 39%) in groups 1 and 2, respectively. Anti-TB medication was given to 7 patients (30%) in group 2 despite the lack of a confirmed diagnosis. The outcome in terms of complications and renal function loss was not significantly different between the 2 groups. Flank pain, renal cavitation, urolithiasis, and ureteral stricture formation were significantly more common in the group with suspected UGTB than in those with confirmed UGTB. However, other clinical characteristics did not differ significantly between the 2 groups. In patients with clinical features highly suspicious of UGTB, it appears reasonable to institute anti-TB treatment, despite the lack of a confirmed diagnosis.


Measurement of vaccination coverage at age 24 and 19-35 months: a case study of multiple imputation in public health.
Childhood immunization coverage in the United States (US) is often measured at age 24 months or, in the National Immunization Survey (NIS) at age of interview, which is between 19 and 35 months. This paper compares these standards. Data from the NIS is used to compare immunization coverage at time of interview, retrospectively among all children aged 24 or more months at time of interview, and obtained via multiple imputation (with 10 imputations) for all children, both nationally, by state, and by demographic groups. At the national level, the difference between the 19-35 month estimate and the 24 month complete-case estimate was 1.9 percentage points. For most but not all states and subgroups, the 19-35 month estimate was higher than the 24 month complete-case estimate. The difference between vaccination coverage measured at 19-35 months and 24 months ranged from -2.3 to 7.5 percentage points among states. For three states, the difference between the 19-35 month and 24 month complete-case estimate was more than 6 percentage points, in twelve states there was a 4-6 percentage point difference, and in sixteen states a 2-4 percentage point difference. Conversely, five states had higher 24 month complete-case estimates than 19-35 month estimates. We found that the coverages at 19-35 and 24 months differ such that they would rarely be adequate surrogates for one another, particularly at a state level. Multiple imputation, which is easily implemented, increases precision of estimates of coverage at age 24 months.


A new model for dermatitis herpetiformis that uses HLA-DQ8 transgenic NOD mice.
Dermatitis herpetiformis (DH) is an autoimmune blistering skin disorder that is associated with gluten sensitivity. It presents as a papulovesicular rash and is often associated with enteropathy. The rash resolves when the patient is placed on a gluten-free diet and/or dapsone. DH, as well as celiac disease, is tightly associated with DQ2 and DQ8. A novel mouse model for DH is described that utilizes the NOD background and the HLA-DQ8 transgene. The addition of DQ8 contributes sensitivity to gliadin, and the addition of the NOD background contributes to autoimmunity and pathogenesis. Fifteen NOD DQ8+ mice of 90 that were sensitized to gluten developed blistering pathology similar to that seen in DH. Neutrophil infiltration of the dermis, deposition of IgA at the dermal-epidermal junction, and a complete reversal of the blistering phenomenon with the administration of a gluten-free diet with or without dapsone were observed. None of the 3 blistering mice examined had small-bowel pathology. This animal model of DH will be useful to determine the specificity of the IgA deposits, as well as the pathogenic mechanisms that occur in the skin as a result of gluten ingestion.


HINT: High-quality protein interactomes and their applications in understanding human disease.
A global map of protein-protein interactions in cellular systems provides key insights into the workings of an organism. A repository of well-validated high-quality protein-protein interactions can be used in both large- and small-scale studies to generate and validate a wide range of functional hypotheses. We develop HINT (http://hint.yulab.org) - a database of high-quality protein-protein interactomes for human, Saccharomyces cerevisiae, Schizosaccharomyces pombe, and Oryza sativa. These were collected from several databases and filtered both systematically and manually to remove low-quality/erroneous interactions. The resulting datasets are classified by type (binary physical interactions vs. co-complex associations) and data source (high-throughput systematic setups vs. literature-curated small-scale experiments). We find strong sociological sampling biases in literature-curated datasets of small-scale interactions. An interactome without such sampling biases was used to understand network properties of human disease-genes - hubs are unlikely to cause disease, but if they do, they usually cause multiple disorders. HINT is of significant interest to researchers in all fields of biology as it addresses the ubiquitous need of having a repository of high-quality protein-protein interactions. These datasets can be utilized to generate specific hypotheses about specific proteins and/or pathways, as well as analyzing global properties of cellular networks. HINT will be regularly updated and all versions will be tracked.


In-season rehabilitation of a division III female wrestler after a glenohumeral dislocation.
Wrestling is a popular sport in the United States at both the high school and collegiate levels. Traditionally a men's sport, participation by female athletes in wrestling is increasing. There exists a paucity of literature regarding injury incidence in women's wrestling. This lack of information challenges the ability of sports medicine and strength training professionals to design optimal injury prevention programs, training routines, and rehabilitation strategies. The objective of this report is to detail the successful conservative rehabilitation of a female wrestler after an initial glenohumeral dislocation. Case report. A 20-year-old female wrestling student-athlete presented to the university's sports medicine team after sustaining an anteriorly dislocated right shoulder. The patient had the goal to return back to competition in time for the National Championships. An evidenced-supported, non-traumatic glenohumeral instability rehabilitation protocol combined with weight-bearing exercises simulating functional sport positions was implemented with the goal of returning the injured collegiate female wrestler back to sport. At the end of the rehabilitation program the athlete demonstrated full active range of motion, good strength in the right shoulder, and reported her pain rating at a 1/10. The conservative rehabilitation strategy utilized in this case enabled the patient to return to wrestling and successfully compete at the National Championships.


Prioritization of cancer marker candidates based on the immunohistochemistry staining images deposited in the human protein atlas.
Cancer marker discovery is an emerging topic in high-throughput quantitative proteomics. However, the omics technology usually generates a long list of marker candidates that requires a labor-intensive filtering process in order to screen for potentially useful markers. Specifically, various parameters, such as the level of overexpression of the marker in the cancer type of interest, which is related to sensitivity, and the specificity of the marker among cancer groups, are the most critical considerations. Protein expression profiling on the basis of immunohistochemistry (IHC) staining images is a technique commonly used during such filtering procedures. To systematically investigate the protein expression in different cancer versus normal tissues and cell types, the Human Protein Atlas is a most comprehensive resource because it includes millions of high-resolution IHC images with expert-curated annotations. To facilitate the filtering of potential biomarker candidates from large-scale omics datasets, in this study we have proposed a scoring approach for quantifying IHC annotation of paired cancerous/normal tissues and cancerous/normal cell types. We have comprehensively calculated the scores of all the 17219 tested antibodies deposited in the Human Protein Atlas based on their accumulated IHC images and obtained 457110 scores covering 20 different types of cancers. Statistical tests demonstrate the ability of the proposed scoring approach to prioritize cancer-specific proteins. Top 100 potential marker candidates were prioritized for the 20 cancer types with statistical significance. In addition, a model study was carried out of 1482 membrane proteins identified from a quantitative comparison of paired cancerous and adjacent normal tissues from patients with colorectal cancer (CRC). The proposed scoring approach demonstrated successful prioritization and identified four CRC markers, including two of the most widely used, namely CEACAM5 and CEACAM6. These results demonstrate the potential of this scoring approach in terms of cancer marker discovery and development. All the calculated scores are available at http://bal.ym.edu.tw/hpa/.


Reappearance of Salmonella serovar Choleraesuis var. Kunzendorf in Danish pig herds.
Salmonella enterica serovar Choleraesuis is a porcine adapted serovar which may cause serious outbreaks in pigs. Here we describe outbreaks of salmonellosis due to S. Choleraesuis in four Danish pig farms in 2012-2013 by clinic, serology, and microbiology and compare the isolates to those of a previous outbreak in 1999-2000. The infection was in some herds associated with high mortality and a moderate to high sero-prevalence was found. In 2012-2013 the disease contributed to increased mortality but occurred concomitant with other disease problems in the herds, which likely delayed the diagnosis by up to several months. Nine isolates from the four farms in 2012-2013 and 14 isolates obtained from the outbreak in Denmark in 1999-2000 were subjected to typing using pulsed-field gel electrophoresis (PFGE). Seven isolates were selected for whole genome sequencing (WGS). The PFGE results of 23 isolates displayed five different profiles. The isolates from 2012 to 2013 revealed two distinct profiles, both different from the isolates recovered in 1999-2000. Two of the 2012-2013 farms shared PFGE profiles and had also transported pigs between them. The profile found in the two other 2012-2013 farms was indistinguishable but no epidemiological connection between these farms was found. Analysis of the number of single nucleotide polymorphisms (SNPs) from the WGS data indicated that the isolates from the farms in 2012-2013 were more closely related to each other than to isolates from the outbreak in 1999. It was therefore concluded that the infection was a new introduction and not a persistent infection since the outbreak in 1999. It may further be suggested that there were two or three independent rather than a single introduction. The re-introduction of S. Choleraesuis in Denmark emphasizes the importance of strict hygiene measures in the herds. Further investigations using WGS are now in progress on a larger collection of isolates to study clonality at European level and trace the origin of the infections.


[Prion protein: structure, functions and polymorphisms associated with human spongiform encephalopathies].
Transmissible subacute spongiform encephalopathies (TSSE), also known as prion diseases, are rare neurodegenerative disorders of both humans and animals. The biochemical hallmark of these diseases is an accumulation in the brain of an abnormal form of the host-encoded prion protein (PrP). This pathological accumulation could result from a protein conformational change under the influence of unknown factors. The normal function of PrP is unknown. The abnormal form is thought to induce neurodegeneration when experimentally or accidentally introduced in recipient hosts. Such a possibility would explain the transmissible character of these diseases illustrated in humans by iatrogenic contamination. Considerable attention has been focused on the host PrP gene and its relation with the genetic susceptibility of humans and animals. Mutations in PRNP, the gene which encodes PrP in humans, are present in 16% of the patients and might be causative. In patients without any PRNP mutation, a coding polymorphism (129 Met/Val) defines a predisposing factor. Since few years, important progress in the molecular genetics of TSSE in both humans and animals have been performed and point out that the development of different forms of these diseases, experimental, iatrogenic or spontaneous, are strongly dependent on the primary structure of the host PrP.


Long-term outcomes and causes of death after acute coronary syndrome in patients in the Bologna, Italy, area.
We sought to evaluate the rates, time course, and causes of death in the long-term follow-up of unselected patients with acute coronary syndromes (ACS). We enrolled 2046 consecutive patients hospitalized from January 2004 to December 2005 with an audited final diagnosis of ACS. The primary study end point was 5-year all-cause mortality. In our series, 896 patients had ST-segment elevation (STE) and 1,150 non-ST-segment elevation (NSTE). Mean age of the study population was 71.6 years. Primary percutaneous coronary intervention was performed in 86% of STE-ACS, and 70% of NSTE-ACS was managed invasively. The 5-year all-cause mortality was 36.4% for STE-ACS and 42.0% for NSTE-ACS, with patients with STE-ACS showing a trend boarding statistical significance toward a lower risk of mortality (hazard ratio [HR] = 0.88, 95% confidence interval [CI] 0.76 to 1.02, p = 0.08). Landmark analysis demonstrated that patients with STE-ACS had a higher risk of 30-day mortality (STE-ACS vs NSTE-ACS HR = 1.53, 95% CI 1.16 to 2.06, p = 0.003) whereas the risk of NSTE-ACS increased markedly after 1 year (STE-ACS vs NSTE-ACS HR = 0.67, 95% CI 0.53 to 0.84, p = 0.001). The contribution of noncardiovascular (CV) causes to overall mortality increased from 3% at 30 days to 34% at 5 years, with cancer and infections being the most common causes of non-CV death both in STE-ACS and NSTE-ACS. In conclusion, long-term mortality after ACS is still too high both for STE-ACS and NSTE-ACS. Although patients with STE-ACS have a higher mortality during the first year, the mortality of patients with NSTE-ACS increases later, when non-CV co-morbidities gain greater importance.


Regulation of lamellipodial persistence, adhesion turnover, and motility in macrophages by focal adhesion kinase.
Macrophages are a key component of the innate immune system. In this study, we investigate how focal adhesion kinase (FAK) and the related kinase Pyk2 integrate adhesion signaling and growth factor receptor signaling to regulate diverse macrophage functions. Primary bone marrow macrophages isolated from mice in which FAK is conditionally deleted from cells of the myeloid lineage exhibited elevated protrusive activity, altered adhesion dynamics, impaired chemotaxis, elevated basal Rac1 activity, and a marked inability to form stable lamellipodia necessary for directional locomotion. The contribution of FAK to macrophage function in vitro was substantiated in vivo by the finding that recruitment of monocytes to sites of inflammation was impaired in the absence of FAK. Decreased Pyk2 expression in primary macrophages also resulted in a diminution of invasive capacity. However, the combined loss of FAK and Pyk2 had no greater effect than the loss of either molecule alone, indicating that both kinases function within the same pathway to promote invasion.


The genetics of complex autoimmune diseases: non-MHC susceptibility genes.
Susceptibility to complex autoimmune diseases (AIDs) is a multigenic phenotype affected by a variety of genetic and environmental or stochastic factors. After over a decade of linkage analyses, the identification of non-major histocompatibility complex (non-MHC) susceptibility alleles has proved to be difficult, predominantly because of extensive genetic heterogeneity and possible epistatic interactions among the multiple genes required for disease development. Despite these difficulties, progress has been made in elucidating the genetic mechanisms that influence the inheritance of susceptibility, and the pace of gene discovery is accelerating. An intriguing new finding has been the colocalization of several AID susceptibility genes in both rodent models and human linkage studies. This may indicate that several susceptibility alleles affect multiple AIDs, or alternatively that genomic organization has resulted in the clustering of many immune system genes. The completion of the human genome sequence, coupled with the imminent completion of the mouse genome, should yield key information that will dramatically enhance the rate of gene discovery in complex conditions such as AID susceptibility.


Global epidemiology of HIV among female sex workers: influence of structural determinants.
Female sex workers (FSWs) bear a disproportionately large burden of HIV infection worldwide. Despite decades of research and programme activity, the epidemiology of HIV and the role that structural determinants have in mitigating or potentiating HIV epidemics and access to care for FSWs is poorly understood. We reviewed available published data for HIV prevalence and incidence, condom use, and structural determinants among this group. Only 87 (43%) of 204 unique studies reviewed explicitly examined structural determinants of HIV. Most studies were from Asia, with few from areas with a heavy burden of HIV such as sub-Saharan Africa, Russia, and eastern Europe. To further explore the potential effect of structural determinants on the course of epidemics, we used a deterministic transmission model to simulate potential HIV infections averted through structural changes in regions with concentrated and generalised epidemics, and high HIV prevalence among FSWs. This modelling suggested that elimination of sexual violence alone could avert 17% of HIV infections in Kenya (95% uncertainty interval [UI] 1-31) and 20% in Canada (95% UI 3-39) through its immediate and sustained effect on non-condom use) among FSWs and their clients in the next decade. In Kenya, scaling up of access to antiretroviral therapy among FSWs and their clients to meet WHO eligibility of a CD4 cell count of less than 500 cells per μL could avert 34% (95% UI 25-42) of infections and even modest coverage of sex worker-led outreach could avert 20% (95% UI 8-36) of infections in the next decade. Decriminalisation of sex work would have the greatest effect on the course of HIV epidemics across all settings, averting 33-46% of HIV infections in the next decade. Multipronged structural and community-led interventions are crucial to increase access to prevention and treatment and to promote human rights for FSWs worldwide.


How can employment-based benefits help the nurse shortage?
During a labor shortage, employment-based benefits can be used to recruit and retain workers. This paper provides data on the availability of benefits to registered nurses (RNs), reports on how health care leaders are approaching the provision of employment-based benefits for nurses, and considers what nurses have to say in focus groups about benefits. Because of the ongoing nurse shortage, many employers are trying to enhance the benefits they offer to support recruitment and retention efforts. We offer recommendations for health care leaders that follow from our findings about the current state of nurses' employment-based benefits.


Knee joint distraction compared with total knee arthroplasty: a randomised controlled trial.
Knee joint distraction (KJD) is a relatively new, knee-joint preserving procedure with the goal of delaying total knee arthroplasty (TKA) in young and middle-aged patients. We present a randomised controlled trial comparing the two. The 60 patients ≤ 65 years with end-stage knee osteoarthritis were randomised to either KJD (n = 20) or TKA (n = 40). Outcomes were assessed at baseline, three, six, nine, and 12 months. In the KJD group, the joint space width (JSW) was radiologically assessed, representing a surrogate marker of cartilage thickness. In total 56 patients completed their allocated treatment (TKA = 36, KJD = 20). All patient reported outcome measures improved significantly over one year (p < 0.02) in both groups. At one year, the TKA group showed a greater improvement in only one of the 16 patient-related outcome measures assessed (p = 0.034). Outcome Measures in Rheumatology-Osteoarthritis Research Society International clinical response was 83% after TKA and 80% after KJD. A total of 12 patients (60%) in the KJD group sustained pin track infections. In the KJD group both mean minimum (0.9 mm, standard deviation (sd) 1.1) and mean JSW (1.2 mm, sd 1.1) increased significantly (p = 0.004 and p = 0.0003). In relatively young patients with end-stage knee osteoarthritis, KJD did not demonstrate inferiority of outcomes at one year when compared with TKA. However, there is a high incidence of pin track infection associated with KJD. Cite this article: Bone Joint J 2017;99-B:51-8.


A link between visual disambiguation and visual memory.
Sensory information in the retinal image is typically too ambiguous to support visual object recognition by itself. Theories of visual disambiguation posit that to disambiguate, and thus interpret, the incoming images, the visual system must integrate the sensory information with previous knowledge of the visual world. However, the underlying neural mechanisms remain unclear. Using functional magnetic resonance imaging (fMRI) of human subjects, we have found evidence for functional specialization for storing disambiguating information in memory versus interpreting incoming ambiguous images. Subjects viewed two-tone, "Mooney" images, which are typically ambiguous when seen for the first time but are quickly disambiguated after viewing the corresponding unambiguous color images. Activity in one set of regions, including a region in the medial parietal cortex previously reported to play a key role in Mooney image disambiguation, closely reflected memory for previously seen color images but not the subsequent disambiguation of Mooney images. A second set of regions, including the superior temporal sulcus, showed the opposite pattern, in that their responses closely reflected the subjects' percepts of the disambiguated Mooney images on a stimulus-to-stimulus basis but not the memory of the corresponding color images. Functional connectivity between the two sets of regions was stronger during those trials in which the disambiguated percept was stronger. This functional interaction between brain regions that specialize in storing disambiguating information in memory versus interpreting incoming ambiguous images may represent a general mechanism by which previous knowledge disambiguates visual sensory information.


Cat admissions to RSPCA shelters in Queensland, Australia: description of cats and risk factors for euthanasia after entry.
A lack of information limits understanding of the excess cat problem and development of effective management strategies. This study describes cats entering Royal Society for the Prevention of Cruelty to Animals (RSPCA) Queensland shelters and identifies risk factors for euthanasia. Data for cats entering relevant shelters (July 2006-June 2008) were obtained from the RSPCA's electronic database. Univariable and multivariable logistic regression analyses were conducted to identify risk factors for euthanasia. Of 33,736 cats admitted, 46% were adult cats (≥3 months) and 54% were kittens (<3 months). The most common reason for admission was stray (54%), followed by owner surrender (44%). Euthanasia was the most common outcome (65%), followed by adoption (30%). The odds of euthanasia were lower for kittens and for cats that were desexed prior to admission. Of the strays, 8% had been desexed. For cats of similar age, sex, desexed and feral status, stray cats were more likely to be adopted than owner-surrenders. Strategies are needed to reduce numbers of cats admitted and euthanased. Given the high proportion of admissions that were kittens, reducing the incidence of delayed sterilisation of owned cats may be an important strategy for reducing the number of unwanted kittens. Many cats admitted as strays were rehomable, but given the high proportion of admissions that are strays, further research on stray populations is needed. Future studies of cats entering shelters would be enhanced if data collection definitions, categories and methods were standardised.


Blood pressure measurement in research and in clinical practice: recent evidence.
For over a century the technique of blood pressure measurement developed by Riva-Rocci and Korotkoff has provided most of the data on hypertension diagnosis and treatment. Its limitations, however, are becoming increasingly evident and therefore alternative solutions are under investigation. This paper is intended to provide an overview of important recent progress in this field, and to highlight future perspectives. A major development in blood pressure measurement is the technical improvement of electronic manometers for use either in the clinic (with the auscultatory approach, as an alternative to use of mercury columns), or in automated oscillometric devices yielding blood pressure measurements devoid of observer-dependency, and allowing long-term blood pressure monitoring. In the latter case, blood pressure measurement is made possible in settings other than the physician's office, either through ambulatory blood pressure monitoring or through self blood pressure measurement at home. These methods are growing in clinical importance, but further studies are needed to define their indications more precisely in the clinical evaluation of hypertensive patients. Recently, important steps towards better standards of blood pressure measurement have been taken, as summarized in the guidelines jointly issued by the European Society of Hypertension (ESH) and the European Society of Cardiology (ESC), in the 7th Joint National Committee Report and (in even more detail) in the Blood Pressure Measurement Guidelines published by the ESH Working Group on Blood Pressure Monitoring. Blood pressure measurement is a rapidly developing field, the importance of which is increasingly acknowledged in the light of the growing awareness of the impact of hypertension on public health. Despite remarkable progress, many methodological issues still remain to be properly addressed.


The free health care initiative: how has it affected health workers in Sierra Leone?
There is an acknowledged gap in the literature on the impact of fee exemption policies on health staff, and, conversely, the implications of staffing for fee exemption. This article draws from five research tools used to analyse changing health worker policies and incentives in post-war Sierra Leone to document the effects of the Free Health Care Initiative (FHCI) of 2010 on health workers.Data were collected through document review (57 documents fully reviewed, published and grey); key informant interviews (23 with government, donors, NGO staff and consultants); analysis of human resource data held by the MoHS; in-depth interviews with health workers (23 doctors, nurses, mid-wives and community health officers); and a health worker survey (312 participants, including all main cadres). The article traces the HR reforms which were triggered by the FHCI and evidence of their effects, which include substantial increases in number and pay (particularly for higher cadres), as well as a reported reduction in absenteeism and attrition, and an increase (at least for some areas, where data is available) in outputs per health worker. The findings highlight how a flagship policy, combined with high profile support and financial and technical resources, can galvanize systemic changes. In this regard, the story of Sierra Leone differs from many countries introducing fee exemptions, where fee exemption has been a stand-alone programme, unconnected to wider health system reforms. The challenge will be sustaining the momentum and the attention to delivering results as the FHCI ceases to be an initiative and becomes just 'business as normal'. The health system in Sierra Leone was fragile and conflict-affected prior to the FHCI and still faces significant challenges, both in human resources for health and more widely, as vividly evidenced by the current Ebola crisis.


Processed Meat Protein and Heat-Stable Peptide Marker Identification Using Microwave-Assisted Tryptic Digestion.
New approaches to rapid examination of proteins and peptides in complex food matrices are of great interest to the community of food scientists. The aim of the study is to examine the influence of microwave irradiation on the acceleration of enzymatic cleavage and enzymatic digestion of denatured proteins in cooked meat of five species (cattle, horse, pig, chicken and turkey) and processed meat products (coarsely minced, smoked, cooked and semi-dried sausages). Severe protein aggregation occurred not only in heated meat under harsh treatment at 190 °C but also in processed meat products. All the protein aggregates were thoroughly hydrolyzed after 1 h of trypsin treatment with short exposure times of 40 and 20 s to microwave irradiation at 138 and 303 W. There were much more missed cleavage sites observed in all microwave-assisted digestions. Despite the incompleteness of microwave-assisted digestion, six unique peptide markers were detected, which allowed unambiguous identification of processed meat derived from the examined species. Although the microwave-assisted tryptic digestion can serve as a tool for rapid and high-throughput protein identification, great caution and pre-evaluation of individual samples is recommended in protein quantitation.


Interaction of the intrinsically unstructured phage lambda N Protein with Escherichia coli NusA.
N protein of the Escherichia coli phage lambda (lambdaN) is involved in antitermination, a transcription regulatory process that is essential for the expression of delayed early genes during phage lytic development. lambdaN is an intrinsically unstructured protein that possesses three distinct binding sites interacting with the carboxy terminus of the E. coli host factor protein NusA, the viral nutBoxB-RNA, and RNA polymerase, respectively. Heteronuclear NMR experiments with lambdaN(1-53) in complex with NusA(339-495) revealed that upon complex formation the lambdaN-binding interface, lambdaN(34-47), adopts a rigid structure. NMR data also indicate the induction of a weak helical structure in the nutboxB RNA-binding region lambdaN(1-22) upon binding to NusA(339-495) even in the absence of RNA. Titration experiments of the lambdaN(1-53)-nutBoxB RNA complex with NusA(339-495) revealed that the ternary complex can be described in terms of two structurally independent binary interactions. Furthermore, chemical-shift perturbation experiments with different NusA constructs and different lambdaN peptides showed that only NusA(353-416) is involved in lambdaN binding. We found that only one molecule of NusA(339-426) binds to one molecule of lambdaN(1-53). We also clarified the role of the lambdaN-binding region and could show that N41-R47 also binds to NusA(339-495). Furthermore, we observe that lambdaN(1-22) adopts a helical fold upon binding to NusA(339-495), in agreement with one of the theoretical models of lambdaN action.


Contributions of mild parkinsonian signs to gait performance in the elderly.
Mild parkinsonian signs (MPS) and gait abnormalities are common in aging, but the association between MPS and objective gait measures is not established in the elderly. This study aims to identify the link between MPS and quantitative gait measures, as well as to determine the pathogenesis of MPS in non-demented community-dwelling older adults without idiopathic Parkinson's disease or other parkinsonian syndromes. Three hundred seventy-four non-demented older adults (mean age, 76.44 ± 6.71 years, 57 % women) participated in this study, where comprehensive neurological and medical assessments were conducted. We defined MPS based on the presence of any one of bradykinesia, rigidity, or rest tremor. Velocity and spatial, temporal, and variability gait parameters were recorded using an instrumented walkway. The associations of MPS and gait parameters as well as the relationship of individual MPS with medical illnesses were assessed with linear regressions controlling for key covariates. Participants with MPS walked slower and with disturbed spatial and variability components of gait compared to those without MPS. Bradykinesia was associated with worse spatial and variability gait parameters. This association was only significant for axial bradykinesia, but not for the presence of bradykinesia in the limbs. Cerebrovascular disease (β = .20, p < .01) was associated with bradykinesia, whereas cardiovascular disease (β = .15, p < .05) was associated with rigidity. Among MPS, bradykinesia but not rigidity or tremor was associated with worse quantitative gait performance in older adults. Cerebrovascular disease, a preventable condition, was specifically associated with bradykinesia.


Role of nonexercise activity thermogenesis (NEAT) in obesity.
Obesity develops when there is an imbalance between energy intake and energy expenditure, which can vary daily within and among individuals. High levels of energy intake and low levels of energy expenditure contribute to obesity, both together and independently. Energy expenditure from exercise associated with formal programs is encouraged for health and weight loss, but most individuals get very little formal exercise. Nonexercise activity thermogenesis (NEAT) is the cumulative energy expended through all other activities of daily living. It is highly variable among individuals; it is controlled by the environment and, possibly, neurobiologically. Mounting evidence suggests that NEAT is critical in determining a person's susceptibility to body fat deposition and is a major factor in human obesity. Recent research supported by the Minnesota Partnership for Biotechnology and Medical Genomics, a state-sponsored collaborative effort of the University of Minnesota and Mayo Clinic, is helping to define the complex brain regulation of NEAT and its role in obesity. This article reviews the evidence for NEAT and the impact of NEAT on obesity.


Importance of adenosine-to-inosine editing adjacent to the anticodon in an Arabidopsis alanine tRNA under environmental stress.
In all organisms, transfer RNAs (tRNAs) undergo extensive post-transcriptional modifications. Although base modifications in the anticodon are known to alter decoding specificity or improve decoding accuracy, much less is known about the functional relevance of modifications in other positions of tRNAs. Here, we report the identification of an A-to-I tRNA editing enzyme that modifies the tRNA-Ala(AGC) in the model plant Arabidopsis thaliana. The enzyme is homologous to Tad1p, a yeast tRNA-specific adenosine deaminase, and it selectively deaminates the adenosine in the position 3'-adjacent to the anticodon (A37) to inosine. We show that the AtTAD1 protein is exclusively localized in the nucleus. The tad1 loss-of-function mutants isolated in Arabidopsis show normal accumulation of the tRNA-Ala(AGC), suggesting that the loss of the I37 modification does not affect tRNA stability. The tad1 knockout mutants display no discernible phenotype under standard growth conditions, but produce less biomass under environmental stress conditions. Our results provide the first evidence in support of a physiological relevance of the A37-to-I modification in eukaryotes.


Tackling inequalities in health: evaluating the New Deal for Communities initiative.
To assess health improvement and differential changes in health across various sociodemographic groups in neighbourhood renewal areas. A longitudinal survey of 10,390 residents in New Deal for Communities (NDC) areas and 977 residents in comparator areas in England. Measures and methods: Changes on several outcomes across five domains (health, unemployment, education, crime and the physical environment) were assessed by sex, age, educational and ethnic group. Small overall improvements were seen on all domains in NDC areas but similar improvements were also seen in comparator areas. In NDC areas, higher educational groups were more likely to stop smoking, less likely to develop a limiting long-term illness, more likely to find employment and more likely to participate in education or training (p for trend <0.05). Older people and women were less likely to find employment and experienced smaller increases in income. These patterns were generally mirrored in comparator areas, although the education gradient in participation in education or training was less steep in NDC areas. Evidence from two-year follow-up does not support an NDC effect, either overall or for particular population groups. Residents with lower education experienced the least favourable health profiles at baseline and the smallest improvements. Programme leaders should consider how to encourage participation among those with the lowest education. A shallower social gradient in participation in education and training in NDC areas and a lack of gradient in income, crime and environmental outcomes indicate that some aspects of the programme may be reaching all sections of the community.


Electrosurgery reduces blood loss and immediate postoperative inflammation compared to cold instruments for midline celiotomy in dogs: A randomized controlled trial.
To compare the use of an electrosurgical device with traditional cold instruments (scalpel and scissors) for midline celiotomy incision. Prospective randomized controlled clinical trial. One hundred and twenty client-owned dogs undergoing abdominal surgery. Dogs were prospectively recruited and randomized to receive electroincision or cold instrument incision. For cold incision, surgeons used basic surgical instruments including scalpel and scissors. For electroincision, surgeons only used the electrosurgical device in cutting mode. Time for the approach, blood loss, and the incision length were recorded. A blinded observer assessed pain and incision redness, swelling, and discharge at 24 and 48 hours postoperative (graded 0-3). Owner assessment of incision healing was recorded by telephone interview. Blood loss during surgery was significantly lower for electroincision (mean 0.7, SD 1.7 mL) than cold incision (mean 3.0, SD 4.3 mL, P < .0001) with no significant difference in incision length or time for approach. Electroincision was associated with significantly less incision redness (cold median 1, range 0-3; electroincision median 0, range 0-2, P = .02) and less incision discharge (cold median 0.5 range 0-3; electroincision median 0, range 0-1, P = .006) at 24 hours postoperative. There was no significant difference in pain scores or incision healing in dogs receiving the two techniques. No incisional hernias were reported. A surgical site infection occurred in 1 dog (cold incision). Electroincision for a celiotomy approach in the dog reduces blood loss, and incision redness and discharge in the immediate postoperative period without affecting the occurrence of wound complications such as infection and dehiscence (including linea alba).


Protein-ribosome-mRNA display: affinity isolation of enzyme-ribosome-mRNA complexes and cDNA cloning in a single-tube reaction.
An enzyme-ribosome-mRNA complex was specifically purified by binding to the immobilized enzyme substrate and the cDNA was cloned in a single-tube reaction by one-step reverse transcription-PCR. The ganglioside GM3, used by sialyltransferase II (ST-II) as a substrate, was coated on a 96-well microtiter plate and ST-II was in vitro transcribed and translated from a cDNA library. The isolation of an enzyme-specific protein-ribosome (PRIME) complex was achieved with as little as 0.1 ng ST-II-specific cDNA in 5 microg of a total plasmid preparation or with the cDNA prepared from sublibraries previously inoculated at a density of 2000 clones/culture well. The affinity purification of the PRIME complex was highly specific for GM3 and did not result in cDNA amplification when a different ganglioside (GM1) was used for coating of the microtiter plate. The amplified cDNA was used for cloning or a second round of ribosome display, providing a fast analysis of enzyme affinity to multiple substrates. PRIME display can be used for host-free cDNA cloning from mRNA or cDNA libraries and for binding site mapping of the in vitro translated protein. The use of a single-tube reaction in ligand-coated microtiter plates indicates the versatility of PRIME display for cDNA cloning by automated procedures.